Recombinant Mouse Coiled-coil domain-containing protein 47 (Ccdc47)

What is CCDC47 and what are its primary cellular functions?

CCDC47 is a calcium-binding endoplasmic reticulum (ER) transmembrane protein that serves multiple essential cellular functions. It functions as a component of the multi-pass translocon (MPT) complex that mediates insertion of multi-pass membrane proteins into the lipid bilayer of membranes . Within this complex, CCDC47 specifically functions in the PAT subcomplex to occlude the lateral gate of the SEC61 complex, which is critical for proper transmembrane protein insertion .

Beyond membrane protein processing, CCDC47 plays crucial roles in:

• Regulation of calcium ion homeostasis in the ER

• Protein degradation via the ERAD (ER-associated degradation) pathway

• Maintenance of ER organization, particularly during embryogenesis

Loss of CCDC47 function results in embryonic lethality in mice, indicating its essential role in early development . Cellular studies demonstrate that CCDC47 deficiency leads to decreased total ER Ca²⁺ storage, impaired Ca²⁺ signaling mediated by the IP₃R Ca²⁺ release channel, and reduced ER Ca²⁺ refilling via store-operated Ca²⁺ entry .

What is the molecular structure and cellular localization of CCDC47?

CCDC47 is an endoplasmic reticulum transmembrane protein containing coiled-coil domains, which are important structural motifs for protein-protein interactions . The protein contains calcium-binding domains with low affinity but high capacity for calcium ions . The mouse CCDC47 protein sequence begins with: MKAFYAFCVVLLVFGSVSEAKFDDFEDEEDIVEYDDNDFAEFEDT and continues through its functional domains .

Regarding cellular localization:

• CCDC47 is primarily localized to the endoplasmic reticulum membrane

• It is positioned to interact with the SEC61 complex at the lateral gate

• As part of the PAT subcomplex, it is strategically positioned to sequester highly polar regions in transmembrane domains away from the non-polar membrane environment

This specific localization is essential for its function in protein insertion, calcium homeostasis, and ER organization during development .

How is the CCDC47 gene expressed and what splice variants exist?

The CCDC47 gene exhibits complex expression patterns. The gene contains 16 distinct gt-ag introns and its transcription produces 9 different mRNAs, consisting of 6 alternatively spliced variants and 3 unspliced forms . The gene structure includes:

• 3 probable alternative promoters

• 3 non-overlapping alternative last exons

• 8 validated alternative polyadenylation sites

The resulting mRNAs differ by:

• Truncation of the 5' end

• Truncation of the 3' end

• Presence or absence of cassette exons

• Overlapping exons with different boundaries

The CCDC47 gene is expressed at high levels in various tissues, with functional implications for calcium ion homeostasis across different cell types . This complex pattern of expression and splicing suggests tissue-specific regulation and potentially specialized functions in different cellular contexts.

What are the optimal methods for expressing and purifying recombinant mouse CCDC47?

For successful expression and purification of recombinant mouse CCDC47, researchers have established effective protocols that maintain protein functionality:

Expression System Selection:
The HEK-293 human embryonic kidney cell line has proven highly effective for expressing mouse CCDC47 protein fragments (Met1-Ser135) with C-terminal His tags . This mammalian expression system provides appropriate post-translational modifications and protein folding environment compared to bacterial systems.

Expression Construct Design:

• DNA sequence encoding mouse Ccdc47 (NP_080285.2) from Met1-Ser135

• Addition of a polyhistidine tag at the C-terminus for purification

• Expression vector with appropriate promoter for high-level expression in HEK-293 cells

Purification Protocol:

• Cell lysis under native conditions preserving protein conformation

• Affinity chromatography using Ni-NTA resin for His-tagged protein capture

• Washing steps to achieve >95% purity as determined by SDS-PAGE

• Quality control to ensure endotoxin levels <1.0 EU per μg (as determined by LAL method)

Storage and Stability:

• Lyophilization from sterile PBS, pH 7.4

• Lyophilized proteins remain stable up to 12 months at -20°C to -80°C

• Reconstituted protein solutions can be stored at 4-8°C for 2-7 days

• Aliquots of reconstituted samples remain stable at <-20°C for 3 months

This methodological approach yields mouse CCDC47 protein with >95% purity and low endotoxin levels, making it suitable for functional studies, antibody production, and protein interaction analyses .

How can researchers effectively study CCDC47's role in calcium homeostasis?

Investigating CCDC47's role in calcium homeostasis requires specialized techniques to measure ER calcium dynamics and signaling:

In Vitro Calcium Imaging Methods:

• ER Ca²⁺ Storage Measurement:

  • Use ER-targeted calcium indicators (e.g., D1ER, G-CEPIA1er)

  • Apply thapsigargin to release ER calcium stores

  • Quantify total releasable calcium pool size in CCDC47-deficient vs. control cells

• IP₃R-Mediated Ca²⁺ Release Assessment:

  • Stimulate cells with IP₃-generating agonists (e.g., ATP, histamine)

  • Record cytosolic calcium transients using ratiometric dyes (Fura-2) or genetically encoded indicators

  • Compare amplitude and kinetics of calcium signals between wild-type and CCDC47-manipulated cells

• Store-Operated Ca²⁺ Entry (SOCE) Analysis:

  • Deplete ER stores with thapsigargin in calcium-free medium

  • Reintroduce extracellular calcium

  • Measure the rate and magnitude of calcium entry

  • Analyze STIM1-Orai1 interaction using FRET or co-immunoprecipitation in CCDC47-deficient backgrounds

Genetic Manipulation Approaches:

• CRISPR/Cas9-mediated CCDC47 knockout or knockdown

• Rescue experiments with wild-type or mutant CCDC47 constructs

• Patient-derived cells harboring CCDC47 variants for validation studies

Protein Interaction Studies:

• Co-immunoprecipitation to identify CCDC47 binding partners in calcium signaling pathways

• Proximity labeling techniques (BioID, APEX) to map the CCDC47 interactome

• In vitro calcium binding assays to characterize direct calcium interactions

These methodological approaches have revealed that CCDC47 deficiency results in decreased total ER Ca²⁺ storage, impaired IP₃R-mediated Ca²⁺ signaling, and reduced SOCE, confirming its essential role in maintaining calcium homeostasis .

What disease phenotypes are associated with CCDC47 dysfunction and how can they be modeled?

CCDC47 dysfunction has been linked to a distinct multisystem disorder with specific clinical manifestations. Researchers can model these conditions using various approaches:

Clinical Phenotype of CCDC47-Associated Disorder:

• Woolly hair

• Liver dysfunction

• Pruritus

• Dysmorphic features

• Hypotonia

• Global developmental delay

• Neurological abnormalities (hyperreflexia, poor head control, non-verbal status)

• Brain imaging abnormalities (minimal prominence of cerebral sulci, ventricular enlargement, global white matter paucity, thin corpus callosum)

Disease Modeling Approaches:

• Patient-Derived Cell Models:

  • Fibroblasts or lymphoblasts from affected individuals

  • iPSC derivation and differentiation into relevant cell types (neurons, hepatocytes)

  • Characterization of cellular phenotypes (ER stress, calcium dysregulation, protein processing defects)

• Animal Models:

  • Complete knockout mice exhibit embryonic lethality with:

    • Delayed development

    • Atrophic neural tubes

    • Heart abnormalities

    • Paucity of blood cells in the dorsal aorta

  • Conditional tissue-specific knockouts to bypass embryonic lethality

  • CRISPR-engineered mice harboring patient-specific mutations

• Molecular Analysis of Patient Mutations:

  • Most pathogenic variants are nonsense or frameshift

  • Variants lead to nonsense-mediated mRNA decay or premature protein truncation

  • Functional consequences include decreased CCDC47 mRNA expression and protein levels

Experimental Readouts for Disease Modeling:

• ER calcium homeostasis disruption

• Impaired protein folding and trafficking

• Activation of ER stress response pathways

• Developmental timing and patterning defects

• Tissue-specific manifestations (neural, hepatic, dermatological)

This multifaceted approach to disease modeling provides insights into the pathophysiological mechanisms of CCDC47-associated disorders and potential therapeutic interventions.

How does CCDC47 function in the multi-pass translocon complex for membrane protein insertion?

CCDC47 plays a specialized role in the complex process of multi-pass membrane protein insertion that can be studied using advanced biochemical and cellular approaches:

CCDC47's Function in Membrane Protein Insertion:

• Sequential Process with SEC61:

  • The SEC61 complex mediates insertion of the first few transmembrane segments

  • The MPT complex (including CCDC47) then takes over for subsequent transmembrane regions

  • CCDC47 specifically occludes the lateral gate of SEC61 to facilitate this process

• Role in the PAT Subcomplex:

  • Within the MPT complex, CCDC47 is part of the PAT subcomplex

  • This subcomplex sequesters highly polar regions in transmembrane domains

  • Protects polar regions from the non-polar membrane environment

  • Maintains them until they can be buried in the interior of the fully assembled protein

Methodological Approaches to Study This Function:

• Reconstitution Systems:

  • In vitro translation systems coupled with ER microsomes

  • Purified components reconstituted in liposomes

  • Cross-linking studies to capture transient interactions during insertion

• Real-time Membrane Insertion Assays:

  • Fluorescence-based reporters to monitor membrane insertion kinetics

  • FRET pairs positioned at key locations to detect conformational changes

  • Single-molecule techniques to visualize insertion events

• Structure-Function Analysis:

  • Mutational analysis of CCDC47 domains involved in SEC61 interaction

  • Cryo-EM studies of the MPT complex with trapped substrates

  • Computational modeling of the dynamic insertion process

• Substrate Specificity Determination:

  • Identification of model substrates dependent on CCDC47

  • Characterization of transmembrane domain features requiring CCDC47

  • Proteomic analysis of membrane proteins affected by CCDC47 deficiency

Understanding CCDC47's precise role in membrane protein insertion has implications for numerous cellular processes dependent on proper membrane protein localization and function, including signaling, transport, and cell-cell communication .

What experimental approaches can address the embryonic requirement for CCDC47?

The embryonic lethality observed in CCDC47-deficient mice indicates its essential developmental role. Researchers can employ several strategies to investigate this function:

Developmental Timing and Tissue-Specific Requirements:

• Conditional and Inducible Knockout Systems:

  • Cre-loxP system for tissue-specific deletion

  • Tamoxifen-inducible CreERT2 for temporal control

  • Tissue-specific promoters to target neural, cardiac, or hepatic tissues

  • Progressive developmental staging of knockout induction to determine critical periods

• Chimeric Analysis:

  • ES cell injection into wild-type blastocysts

  • Tracking the contribution of CCDC47-deficient cells to different tissues

  • Competition assays between wild-type and mutant cells

• Developmental Phenotype Characterization:

  • Detailed embryonic staging analysis

  • Histological assessment of:

    • Neural tube formation

    • Cardiac development

    • Hematopoietic cell emergence in the dorsal aorta

    • Early liver bud formation

Molecular Mechanisms Underlying Developmental Requirements:

• Transcriptomic Analysis:

  • RNA-seq of early embryonic tissues in conditional knockouts

  • Single-cell RNA-seq to identify cell populations most affected

  • Integration with developmental trajectory analysis

• Calcium Signaling in Development:

  • Live calcium imaging in developing embryos using genetically encoded indicators

  • Correlation of calcium transients with morphogenetic movements

  • Rescue experiments with calcium ionophores or targeted calcium modulators

• ER Stress and UPR in Development:

  • Analysis of ER stress markers during embryogenesis

  • Assessment of unfolded protein response (UPR) activation

  • Connection between ER homeostasis and developmental patterning

• Mouse Embryonic Fibroblast (MEF) Studies:

  • Isolation of MEFs from CCDC47-deficient embryos before lethality

  • Characterization of cellular phenotypes

  • Comparison with patient-derived cells harboring hypomorphic alleles

These approaches would provide comprehensive insights into why CCDC47 is essential for embryonic development and how its absence leads to developmental failure, with potential implications for understanding human developmental disorders.

What are the quality control parameters for recombinant mouse CCDC47 protein preparations?

Ensuring the quality and functionality of recombinant mouse CCDC47 protein is critical for reliable experimental outcomes. Researchers should implement comprehensive quality control measures:

Purity Assessment:

• SDS-PAGE analysis showing >95% purity

• Absence of degradation products or contaminant bands

• Mass spectrometry confirmation of intact protein

Endotoxin Testing:

• Limulus Amebocyte Lysate (LAL) testing

• Acceptable endotoxin levels <1.0 EU per μg of protein

• Critical for cell-based assays to prevent inflammatory responses

Protein Concentration Determination:

• Bradford or BCA protein assays

• UV spectroscopy (A280)

• Comparison against BSA standards

Functional Verification:

• Calcium binding capacity assessment

• Circular dichroism to confirm proper secondary structure

• Thermal shift assays to evaluate protein stability

Storage Stability Testing:

• Accelerated stability studies at different temperatures

• Freeze-thaw cycle tolerance evaluation

• Long-term stability monitoring

Batch-to-Batch Consistency:

• Lot comparison by SDS-PAGE and functional assays

• Certificate of analysis for each production batch

• Reference standards for inter-batch calibration

The commercial preparations of recombinant mouse CCDC47 (fragment Met1-Ser135) have been validated to meet these rigorous quality control parameters, making them suitable for a wide range of applications including structural studies, functional assays, and antibody production .

How can researchers investigate CCDC47's role in the ERAD pathway?

The endoplasmic reticulum-associated degradation (ERAD) pathway is critical for removing misfolded proteins, and CCDC47 plays a required role in this process. Researchers can employ several methodological approaches:

ERAD Substrate Degradation Assays:

• Pulse-Chase Analysis:

  • Radioactive labeling of newly synthesized proteins

  • Immunoprecipitation of specific ERAD substrates

  • Quantification of degradation rates in CCDC47-deficient vs. control cells

• Fluorescent Timer Proteins:

  • Expression of fluorescent ERAD substrates that change color with time

  • Live-cell imaging to track degradation kinetics

  • Flow cytometry for high-throughput analysis

ERAD Component Interactions:

• Co-immunoprecipitation Studies:

  • Pull-down of CCDC47 to identify interacting ERAD machinery components

  • Reciprocal IPs with known ERAD components (HRD1, SEL1L, OS-9)

  • Western blot analysis of complex formation

• Proximity Labeling Approaches:

  • BioID or TurboID fusion with CCDC47

  • Identification of proximal proteins in the ERAD pathway

  • Mass spectrometry analysis of the biotinylated proteome

Ubiquitination and Retrotranslocation:

• Ubiquitination Assays:

  • Detection of ubiquitinated ERAD substrates

  • Analysis of ubiquitination patterns in CCDC47-deficient cells

  • In vitro ubiquitination reactions with purified components

• Retrotranslocation Monitoring:

  • Split fluorescent protein reporters spanning the ER membrane

  • Protease protection assays to detect cytosolic exposure

  • Cell fractionation to quantify substrate localization

Proteasomal Degradation Analysis:

• Proteasome Inhibition Studies:

  • Treatment with MG132 or bortezomib

  • Accumulation patterns of ERAD substrates

  • Comparison between wild-type and CCDC47-deficient conditions

• Cellular Stress Responses:

  • Analysis of UPR markers (XBP1 splicing, ATF6 cleavage, PERK phosphorylation)

  • ER stress sensor activation in response to ERAD impairment

  • Correlation with cell viability and apoptosis induction

By implementing these methodological approaches, researchers can elucidate CCDC47's specific contributions to the ERAD pathway and how its dysfunction may lead to accumulation of misfolded proteins and subsequent cellular stress.

What analytical methods are most effective for studying CCDC47 in developmental contexts?

Investigating CCDC47's developmental roles requires specialized analytical methods spanning molecular, cellular, and organismal levels:

Developmental Expression Analysis:

• Spatiotemporal Expression Profiling:

  • In situ hybridization to localize CCDC47 mRNA in embryonic tissues

  • Immunohistochemistry for protein distribution during development

  • Reporter gene constructs (e.g., CCDC47-GFP) for live imaging

• Single-Cell Approaches:

  • scRNA-seq to identify cell populations expressing CCDC47

  • Trajectory analysis to correlate expression with developmental fate decisions

  • Integration with developmental atlases

Embryonic Phenotyping Methods:

• Advanced Imaging Techniques:

  • Optical projection tomography for whole-embryo analysis

  • Light sheet microscopy for 3D visualization of developing structures

  • Intravital imaging to track developmental processes in real-time

• Functional Assessment:

  • Embryonic ECG for cardiac function evaluation

  • Neural tube closure quantification

  • Morphometric analysis of anatomical structures

Molecular Developmental Mechanisms:

• Differential Proteomics:

  • TMT or iTRAQ labeling for quantitative comparison

  • Analysis of protein expression changes in CCDC47-deficient embryos

  • Focused analysis of calcium-dependent developmental pathways

• Epigenetic Profiling:

  • ATAC-seq to assess chromatin accessibility changes

  • ChIP-seq for histone modifications affected by calcium signaling

  • Integration with transcriptomic data

• Interactome Analysis in Developmental Contexts:

  • Stage-specific pull-downs to identify developmental binding partners

  • Comparison of interactions across different embryonic tissues

  • Validation using genetic interaction studies

Calcium Signaling in Development:

• Calcium Imaging in Embryonic Tissues:

  • GCaMP sensors for calcium transient visualization

  • Correlation with morphogenetic movements

  • Pharmacological manipulation of calcium signaling

• ER Calcium Dynamics:

  • ER-targeted calcium indicators in developing tissues

  • Analysis of calcium store content during critical developmental windows

  • Connection between calcium homeostasis and developmental milestones

These analytical methods provide a comprehensive toolkit for investigating CCDC47's essential role in embryonic development and the cellular mechanisms through which it influences developmental processes.

What are the potential therapeutic implications of CCDC47 research?

Research on CCDC47 has revealed its crucial roles in calcium homeostasis, protein processing, and development, suggesting several therapeutic avenues:

Targeting Calcium Signaling Pathways:

• Small Molecule Modulators:

  • Compounds that normalize calcium flux in CCDC47-deficient cells

  • Selective ER calcium channel modulators

  • Store-operated calcium entry enhancers

• Gene Therapy Approaches:

  • AAV-mediated delivery of functional CCDC47

  • CRISPR-based correction of pathogenic variants

  • Targeted activation of compensatory calcium regulatory mechanisms

Addressing ER Stress and Protein Misfolding:

• Chemical Chaperones:

  • 4-PBA or TUDCA to alleviate ER stress

  • Selective reduction of misfolded protein burden

  • Prevention of UPR-mediated cellular dysfunction

• Proteasome and ERAD Modulators:

  • Enhancers of residual ERAD function

  • Selective inhibitors of specific ERAD branches

  • Tissue-specific targeting strategies

Developmental Disorder Interventions:

• Critical Developmental Windows:

  • Identification of key periods for intervention

  • Tissue-specific approaches targeting most affected systems

  • Preventive strategies during embryonic development

• Symptom-Specific Treatments:

  • Management of neurological manifestations

  • Approaches for hepatic dysfunction

  • Targeted interventions for skin and hair abnormalities

Biomarker Development:

• Diagnostic Markers:

  • Calcium flux assays in patient cells

  • ER stress signatures

  • Developmental monitoring parameters

• Treatment Response Indicators:

  • Real-time monitoring of calcium homeostasis

  • Quantitative assessment of ER function

  • Developmental milestone achievement metrics

Research into these therapeutic avenues requires detailed understanding of CCDC47's molecular functions and the downstream consequences of its dysfunction in different cellular contexts and developmental stages.

How can large-scale genomic and proteomic analyses advance CCDC47 research?

Leveraging advanced -omics technologies can significantly enhance our understanding of CCDC47 biology:

Genomic Approaches:

• Population Genomics:

  • Analysis of CCDC47 variants across different populations

  • Identification of hypomorphic alleles with partial function

  • Correlation between variant types and phenotypic severity

• Functional Genomics:

  • CRISPR screens for genetic interactions with CCDC47

  • Synthetic lethality mapping

  • Enhancer and suppressor identification

Transcriptomic Analyses:

• RNA-Seq Applications:

  • Differential gene expression in CCDC47-deficient models

  • Alternative splicing changes

  • Non-coding RNA regulation

• Ribosome Profiling:

  • Translation efficiency changes in CCDC47-deficient cells

  • Identification of translationally regulated targets

  • ER-specific translation dynamics

Proteomic Investigations:

• Global Proteome Analysis:

  • Quantitative proteomics of CCDC47-deficient systems

  • Post-translational modification profiling

  • Protein stability and turnover assessment

• Interactome Mapping:

  • Comprehensive CCDC47 interaction networks

  • Dynamic changes during development or stress

  • Tissue-specific interactors

• Spatial Proteomics:

  • Subcellular localization changes in CCDC47-deficient cells

  • Proximity labeling to map microenvironments

  • Correlative microscopy with proteomic analysis

Integrative Multi-Omics:

• Data Integration Strategies:

  • Combined analysis of genomic, transcriptomic, and proteomic datasets

  • Network analysis to identify central nodes and pathways

  • Machine learning approaches to predict functional relationships

• Systems Biology Modeling:

  • Mathematical modeling of calcium homeostasis

  • Prediction of cellular responses to CCDC47 perturbation

  • Integration with developmental timing models

These large-scale approaches can reveal previously unrecognized functions of CCDC47, identify novel therapeutic targets, and place CCDC47 in a broader biological context of cellular homeostasis and development.

What are the emerging techniques for studying CCDC47's structural biology and interaction network?

Recent advances in structural biology and interaction proteomics offer new opportunities to understand CCDC47 at the molecular level:

Structural Biology Approaches:

• Cryo-Electron Microscopy:

  • High-resolution structures of CCDC47 alone or in complexes

  • Visualization of conformational changes upon calcium binding

  • Structure of CCDC47 within the PAT subcomplex of the MPT

• Integrative Structural Biology:

  • Combination of X-ray crystallography for domain structures

  • NMR for dynamic regions

  • Cross-linking mass spectrometry for interface mapping

  • Molecular dynamics simulations to model conformational changes

• AlphaFold2 and Structure Prediction:

  • In silico modeling of full-length CCDC47

  • Prediction of interaction interfaces

  • Analysis of pathogenic variant impacts on structure

Protein-Protein Interaction Technologies:

• Proximity-Based Methods:

  • BioID, TurboID, or APEX2 fusion with CCDC47

  • Identification of transient and stable interactors

  • Organelle-specific interaction mapping

• Advanced Co-IP Approaches:

  • Quantitative SILAC-based interaction proteomics

  • Cross-linking before immunoprecipitation to capture weak interactions

  • Native protein complex isolation followed by mass spectrometry

• Protein Complementation Assays:

  • Split luciferase or fluorescent protein systems

  • Live-cell monitoring of interactions

  • High-throughput screening platforms

Functional Interaction Analysis:

• Genetic Interaction Mapping:

  • CRISPR interference or activation screens

  • Double knockout/knockdown studies

  • Rescue experiments with domain-specific mutants

• Single-Molecule Methods:

  • FRET-based assays for protein-protein interactions

  • Single-molecule tracking in live cells

  • Super-resolution microscopy to visualize CCDC47 in the ER membrane

• Protein Engineering Approaches:

  • Domain swapping to identify functional modules

  • Minimal functional constructs

  • Synthetic binding partners for functional perturbation

These emerging techniques will provide unprecedented insights into CCDC47's structural organization, dynamic interactions, and molecular mechanisms, advancing our understanding of its role in health and disease.

What statistical approaches are recommended for analyzing data from CCDC47 functional studies?

Experimental Design Considerations:

• Power Analysis:

  • Sample size determination based on expected effect sizes

  • Consideration of biological variability in CCDC47 expression

  • Stratification based on genotype or treatment groups

• Randomization and Blinding:

  • Randomized assignment to experimental groups

  • Blinded analysis of phenotypic outcomes

  • Consideration of batch effects in protein preparation

Statistical Methods for Different Data Types:

• Calcium Imaging Data:

  • Area under curve (AUC) measurements for calcium transients

  • Peak amplitude and decay rate comparisons

  • Mixed-effects models for repeated measures

  • Time series analysis for oscillatory patterns

• Protein Expression Analysis:

  • Normalization strategies for western blots

  • ANOVA with post-hoc tests for multiple group comparisons

  • Regression analysis for dose-response relationships

  • Bootstrapping for robust confidence intervals

• Cell-Based Functional Assays:

  • Survival analysis for time-to-event data

  • Multi-parametric analysis for high-content imaging

  • Principal component analysis for dimensional reduction

  • Machine learning classification of cellular phenotypes

Multiple Testing Correction:

• False Discovery Rate Control:

  • Benjamini-Hochberg procedure for large-scale comparisons

  • q-value calculation and reporting

  • Conservative thresholds for exploratory analyses

• Family-wise Error Rate Control:

  • Bonferroni or Šidák correction for confirmatory testing

  • Dunnett's test for comparisons against a control

  • Tukey's HSD for all pairwise comparisons

Reporting and Visualization:

These statistical approaches ensure robust analysis of CCDC47 functional data, facilitating reproducibility and valid interpretation of experimental findings across different research contexts.

Comparative protein characteristics of mouse CCDC47 and its human ortholog

Known CCDC47 pathogenic variants and their functional consequences